The overall objective of this grant is to perform preclinical studies that are needed to successfully express and test HIV and malaria antigens in an attenuated mycobacterium tuberculosis (MTB) vector. Bill Jacobs has developed two double deleted mutant strains of attenuated MTB (delta RD1, delta panCD and delta lysA, delta panCD) that are safer in immunocompromised mice than BCG and protect against MTB challenge as well as BCG. Using these attenuated strains of MTV, we ultimately aim to develop a trivalent vaccine for HIV, tuberculosis (TB), and malaria in attenuated MTB. Specific objectives include: (1) Test attenuated strains of MTB for their virulence in mice and cynomolgous monkeys, and test for their ability to protect cynomolgous monkeys against challenge with MTB (Project 1); (2) Test attenuated MTB MSP-1[42] malaria construct for safety, and test the attenuated MTB-MSP-1[19] and MSP-1[42] vectors for their ability to protect against P. knowlesi challenge in rhesus monkeys (Project 2) as proof of principle; (3) Test BCG and attenuated MTB as vectors for expression of HIV antigens of African HIV isolates that induce broadly reactive MHC Class I restricted CTL in mice and rhesus monkeys (Project 3); (4) Test BCG and attenuated MTB as vectors for expression of HIV envelope constructs that induce broadly reactive neutralizing antibodies to African HIV primary isolates (Project 4); (5) Perform "research with service" and studies of host response gene polymorphorisms in Lusaka, Zambia and in Moshi, Tanzania to begin Phase I vaccine trial preparedness in these communities (Project 5); and (6) Perform GMP preparation of clinical trial immunogens or safety and toxicity studies for attenuated MTB expressing malaria and HIV proteins, and take the first candidates to IND submission to the FDA for approval for Phase 1 safety, toxicity and immunogenicity testing, first in the US, and then in Africa (GMP Facility Core B). Taken together, these studies will perform the research needed to determine the feasibility of attenuated MTB expressing HIV and malaria genes as experimental HIV, TB, and malaria vaccines.